LOCUS       sequence-387444-        5290 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encoding guide RNAs for the knock out of TPS1 and GSY2 genes.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5290)
  AUTHORS   Takhaveev V, Ozsezen S, Smith EN, Zylstra A, Chaillet ML, Chen H, 
            Papagiannakis A, Milias-Argeitis A, Heinemann M
  TITLE     Temporal segregation of biosynthetic processes is responsible for 
            metabolic oscillations during the budding yeast cell cycle.
  JOURNAL   Nat Metab. 2023 Feb;5(2):294-313. doi: 10.1038/s42255-023-00741-x. 
            Epub 2023 Feb 27.
  PUBMED    36849832
REFERENCE   2  (bases 1 to 5290)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1038/s42255-023-00741-x"; journalName: "Nat Metab"; date: 
            "2023-02"; volume: "5"; issue: "2"; pages: "294-313"
COMMENT     Sequence Label: pROS_phleo-Tps1/Gsy2
FEATURES             Location/Qualifiers
     source          1..5290
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(351..368)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(522..1110)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(602..621)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1281..2072,2073..2141))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     1904..1923
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2142..2246)
                     /gene="bla"
                     /label=AmpR promoter
     terminator      complement(2393..2590)
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     promoter        complement(2983..3326)
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     promoter        3481..3749
                     /label=SNR52 promoter
                     /note="promoter for the S. cerevisiae small nucleolar RNA 
                     gene SNR52"
     misc_RNA        3770..3844
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     terminator      3849..3868
                     /label=SUP4 terminator
                     /note="transcription terminator for the S. cerevisiae SUP4 
                     tRNA gene"
     rep_origin      4093..5074
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      complement(5120..5139)
                     /label=SUP4 terminator
                     /note="transcription terminator for the S. cerevisiae SUP4 
                     tRNA gene"
     misc_RNA        complement(5144..5218)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        complement(join(5239..5290,1..218))
                     /label=SNR52 promoter
                     /note="promoter for the S. cerevisiae small nucleolar RNA 
                     gene SNR52"
ORIGIN
        1 caactttcac ttctacagcg tttgaccaaa atcttttgaa cagaacattg tagggtgtga
       61 aaaaatgcgc acctttaccg ctagcccaag agggcactac aaaatctaga gttgtacttc
      121 aaacgtacat gtaatcacct tgtatatact cgaaagaaaa catcaagttt ctgtataaat
      181 atgagtgaaa gcataatcat acattatctt ttcaaagaag ctccagcttt tgttcccttt
      241 agtgagggta ttcacgtaga cggataggta tagccagaca tcagcagcat acttcgggaa
      301 ccgtaggctc tagagagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
      361 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
      421 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
      481 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
      541 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
      601 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
      661 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
      721 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
      781 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
      841 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
      901 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
      961 aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     1021 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     1081 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     1141 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     1201 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     1261 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     1321 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     1381 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     1441 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     1501 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     1561 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     1621 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     1681 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     1741 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     1801 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     1861 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     1921 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     1981 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     2041 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     2101 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     2161 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     2221 aaaaataaac aaataggggt tccgcgcact cgagactata tgtgaaggca tggctatggc
     2281 acggcagaca ttccgccaga tcatcaatag gcaccttcgt acgctgcagg attaagggtt
     2341 ctcgagagct cgttttcgac actggatggc ggcgttagta tcgaatcgac agcagtatag
     2401 cgaccagcat tcacatacga ttgacgcatg atattacttt ctgcgcactt aacttcgcat
     2461 ctgggcagat gatgtcgagg cgaaaaaaaa tataaatcac gctaacattt gattaaaata
     2521 gaacaactac aatataaaaa aactatacaa atgacaagtt cttgaaaaca agaatctttt
     2581 tattgtcagt actgatcatg agatgcctgc aagcaattcg ttctgtatca ggcgcaggag
     2641 cgtcccgtcc gggtcgacca aagcggccat cgtgcctccc cactcctgca gttcgggggc
     2701 atggatgcgc ggatagccgc tgctggtttc ctggatgccg acggatttgc actgccggta
     2761 gaactccgcg aggtcgtcca gcctcaggca gcagctgaac caactcgcga ggggatcgag
     2821 cccggggtgg gcgaagaact ccagcatgag atccccgcgc tggaggatca tccagccggc
     2881 gtcccggaaa acgattccga agcccaacct ttcatagaag gcggcggtgg gatcgaaatc
     2941 tcgtgatggc aggttgggcg tcgcttggtc ggtcataccc atggttgttt atgttcggat
     3001 gtgatgtgag aactgtatcc tagcaagatt ttaaaaggaa gtatatgaaa gaagaacctc
     3061 agtggcaaat cctaaccttt tatatttctc tacaggggcg cggcgtgggg acaattcaac
     3121 gcgtctgtga ggggagcgtt tccctgctcg caggtctgca gcgaggagcc gtaatttttg
     3181 cttcgcgccg tgcggccatc aaaatgtatg gatgcaaatg attatacatg gggatgtatg
     3241 ggctaaatgt acgggcgaca gtcacatcat gcccctgagc tgcgcacgtc aagactgtca
     3301 aggagggtat tctgggcctc catgtcgctg gccgggtgac ccggcgggga cgaggcaagc
     3361 taaacagatc tctagaccta tggcctatgc cacctttcga gaggacgatg cccgtgtcta
     3421 aatgattcga ccagcctaag aatgttcaac ccctcactaa agggaacaaa agctggagct
     3481 tctttgaaaa gataatgtat gattatgctt tcactcatat ttatacagaa acttgatgtt
     3541 ttctttcgag tatatacaag gtgattacat gtacgtttga agtacaactc tagattttgt
     3601 agtgccctct tgggctagcg gtaaaggtgc gcattttttc acaccctaca atgttctgtt
     3661 caaaagattt tggtcaaacg ctgtagaagt gaaagttggt gcgcatgttt cggcgttcga
     3721 aacttctccg cagtgaaaga taaatgatca atttgtaaaa aagacaagag ttttagagct
     3781 agaaatagca agttaaaata aggctagtcc gttatcaact tgaaaaagtg gcaccgagtc
     3841 ggtggtgctt tttttgtttt ttatgtcttc gagtcatgta attagttatg tcacgcttac
     3901 gttcacgccc tccacgcatt taagcataaa cacgcactat gccgttcttc tcatgtatat
     3961 atatatacag gcaacacgca gatataggtg cgacgtgaac agtgagctgt atgtgcgcag
     4021 ctcgcgttgc attttcggaa gcgctcgttt tcggaaacgc tttgaagttc ctattccgaa
     4081 gttcctattc tgtagaaagt ataggaacct cagagcgctt ttgaaaacca aaagcgctct
     4141 gaagacgcac tttcaaaaaa ccaaaaacgc accggactgt aacgagctac taaaatattg
     4201 cgaataccgc ttccacaaac attgctcaaa agtatctctt tgctatatat ctctgtgcta
     4261 tatccctata taacctaccc atccaccttt cgctccttga acttgcatct aaactcgacc
     4321 tctacatttt ttatgtttat ctctagtatt actctttaga caaaaaaatt gtagtaagaa
     4381 ctattcatag agtgaatcga aaacaatacg aaaatgtaaa catttcctat acgtagtata
     4441 tagagacaaa atagaagaaa ccgttcataa ttttctgacc aatgaagaat catcaacgct
     4501 atcactttct gttcacaaag tatgcgcaat ccacatcggt atagaatata atcggggatg
     4561 cctttatctt gaaaaaatgc acccgcagct tcgctagtaa tcagtaaacg cgggaagtgg
     4621 agtcaggctt tttttatgga agagaaaata gacaccaaag tagccttctt ctaaccttaa
     4681 cggacctaca gtgcaaaaag ttatcaagag actgcattat agagcgcaca aaggagaaaa
     4741 aaagtaatct aagatgcttt gttagaaaaa tagcgctctc gggatgcatt tttgtagaac
     4801 aaaaaagaag tatagattct ttgttggtaa aatagcgctc tcgcgttgca tttctgttct
     4861 gtaaaaatgc agctcagatt ctttgtttga aaaattagcg ctctcgcgtt gcatttttgt
     4921 tttacaaaaa tgaagcacag attcttcgtt ggtaaaatag cgctttcgcg ttgcatttct
     4981 gttctgtaaa aatgcagctc agattctttg tttgaaaaat tagcgctctc gcgttgcatt
     5041 tttgttctac aaaatgaagc acagatgctt cgttggaggg cgtgaacgta agcgtgacat
     5101 aactaattac atgactcgaa gacataaaaa acaaaaaaag caccaccgac tcggtgccac
     5161 tttttcaagt tgataacgga ctagccttat tttaacttgc tatttctagc tctaaaactg
     5221 aatggtgcta ttattgtaga tcatttatct tttcactgcg gagaagtttc gaacgccgaa
     5281 acatgcgcac
//
